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The formamidase (FMD) enzyme plays an important role in fungal thriving by releasing a secondary nitrogen source as a product of its activity. In Paracoccidioides species, previous studies have demonstrated the upregulation of this enzyme in a wide range of starvation and infective-like conditions. However, Paracoccidioides lutzii formamidase has not yet been defined as a virulence factor. Here, by employing in vivo infections using an fmd-silenced strain in Galleria mellonella larvae model, we demonstrate the influence of formamidase in P. lutzii's immune stimulation and pathogenicity. The formamidase silencing resulted in improper arrangement of the nodules, poor melanogenesis and decreased fungal burden. Thus, we suggest that formamidase may be a piece composing the process of molecular recognition by Galleria immune cells. Furthermore, formamidase silencing doubled the observed survival rate of the larvae, demonstrating its importance in fungal virulence in vivo. Therefore, our findings indicate that formamidase contributes to Galleria's immune incitement and establishes the role of this enzyme as a P. lutzii virulence factor. Author summary: The study of fungal neglected pathogens is of significant importance for the elucidation of the mechanisms underlying the diseases they cause. The Paracoccidioides lutzii fungus is a pathogen endemic to Latin America, which causes disease in vulnerable portions of society. To cause the disease, the fungal cells must overcome the immune system and obtain nutrients that are typically withdrawn by the host. In this regard, different molecules are crucial for the establishment of infection. The formamidase enzyme plays a role in the survival of the fungus P. lutzii. Our study, presented below, demonstrates that formamidase silencing affects the immune response of G. mellonella larvae to the fungus, resulting in impairment of nodule formation, diminished melanin production, and a reduction in fungal burden. Furthermore, silencing formamidase expression in P. lutzii doubles the survival rate of larvae infected by the fungus, showing its importance for these yeast cells to infect this animals. [ABSTRACT FROM AUTHOR]

Postharvest rot caused by various fungal pathogens is a damaging disease affecting kiwifruit production and quality, resulting in significant annual economic losses. This study focused on isolating the strain P3-1W, identified as Diaporthe eres, as the causal agent of 'Hongyang' postharvest rot disease in China. The investigation highlighted cell wall degrading enzymes (CWDEs) as crucial pathogenic factors. Specially, the enzymatic activities of cellulase, β-galactosidase, polygalacturonase, and pectin methylesterases peaked significantly on the second day after infection of D. eres P3-1W. To gain a comprehensive understanding of these CWDEs, the genome of this strain was sequenced using PacBio and Illumina sequencing technologies. The analysis revealed that the genome of D. eres P3-1W spans 58,489,835 bp, with an N50 of 5,939,879 bp and a GC content of 50.7%. A total of 15,407 total protein-coding genes (PCGs) were predicted and functionally annotated. Notably, 857 carbohydrate-active enzymes (CAZymes) were identified in D. eres P3-1W, with 521 CWDEs consisting of 374 glycoside hydrolases (GHs), 108 carbohydrate esterase (CEs) and 91 polysaccharide lyases (PLs). Additionally, 221 auxiliary activities (AAs), 91 glycosyltransferases (GTs), and 108 carbohydrate binding modules (CBMs) were detected. These findings offer valuable insights into the CAZymes of D. eres P3-1W. [ABSTRACT FROM AUTHOR]

Heat shock proteins are constitutively expressed chaperones induced by cellular stress, such as changes in temperature, pH, and osmolarity. These proteins, present in all organisms, are highly conserved and are recruited for the assembly of protein complexes, transport, and compartmentalization of molecules. In fungi, these proteins are related to their adaptation to the environment, their evolutionary success in acquiring new hosts, and regulation of virulence and resistance factors. These characteristics are interesting for assessment of the host adaptability and ecological transitions, given the emergence of infections by these microorganisms. Based on phylogenetic inferences, we compared the sequences of HSP9, HSP12, HSP30, HSP40, HSP70, HSP90, and HSP110 to elucidate the evolutionary relationships of different fungal organisms to suggest evolutionary patterns employing the maximum likelihood method. By the different reconstructions, our inference supports the hypothesis that these classes of proteins are associated with pathogenic gains against endothermic hosts, as well as adaptations for phytopathogenic fungi. [ABSTRACT FROM AUTHOR]

Ferulic acid (FA), also known as hydroxycinnamic acid, is a sweet-smelling and natural antioxidant present in the cell walls of plants and bran of cereals, making it a common dietary component. FA possesses diverse medicinal properties, including anti-inflammatory, anti-arrhythmic, antioxidant, anti-bacterial, anti-thrombotic, anti-cancer, anti-diabetic, anti-cardiovascular, neuroprotective, anti-apoptotic, antifibrotic, anti-platelet, anti-aging, anti-melanogenesis, angiogenesis promoting, and skin damage reducing effects. Judging by these outstanding credentials of FA, this study investigated the hepatoprotective, spermato-protective, and hemato-protective effects of FA against 2MEETH-induced spermatotoxicity, hematotoxicity, and hepatotoxicity in rats. After oral administration of 2MEETH for 30 days, the number of abnormal and dead spermatozoa, as well as serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and activity of alcohol dehydrogenase (ADH), were significantly increased. In contrast, the number of normal and live spermatozoa, sperm motility and concentration, serum total cholesterol level, red blood cell (RBC), packed cell volume (PCV), and white blood cell (WBC) counts were significantly decreased compared to the control group. FA treatments did not have any effect on all the sperm and hematological parameters checked, but significantly lowered the serum levels of AST, ALT, and ADH activity compared to rats administered with only 2MEETH. Based on the results, it is concluded that FA may not possess spermato- and hemato-protective effects against 2MEETH-induced spermatotoxicity and hematotoxicity but may possess a hepatoprotective effect in rats. [ABSTRACT FROM AUTHOR]

Nowadays, dermatophyte infections are relatively easy to cure, especially since the introduction of orally administered antifungals such as terbinafine and itraconazole. However, these drugs may cause side effects due to liver damage or their interactions with other therapeutics. Hence, the search for new effective chemotherapeutics showing antidermatophyte activity seems to be the urge of the moment. Potassium salts of N-acylhydrazinecarbodithioates are used commonly as precursors for the synthesis of biologically active compounds. Keeping that in mind, the activity of a series of five potassium N-acylhydrazinecarbodithioates (1a–e) and their aminotriazole-thione derivatives (2a–e) was evaluated against a set of pathogenic, keratinolytic fungi, such as Trichophyton ssp., Microsporum ssp. and Chrysosporium keratinophilum, but also against some Gram-positive and Gram-negative bacteria. All tested compounds were found non-toxic for L-929 and HeLa cells, with the IC30 and IC50 values assessed in the MTT assay above 128 mg/L. The compound 5-amino-3-(naphtalene-1-yl)-4,5-dihydro-1H-1,2,4-triazole-5-thione (2d) was found active against all fungal strains tested. Scanning Electron Microscopy (SEM) revealed inhibition of mycelium development of Trichophyton rubrum cultivated on nail fragments and treated with 2d 24 h after infection with fungal spores. Transmission Electron Microscopy (TEM) observation of mycelium treated with 2d showed ultrastructural changes in the morphology of germinated spores. Finally, the RNA-seq analysis indicated that a broad spectrum of genes responded to stress induced by the 2d compound. In conclusion, the results confirm the potential of N-acylhydrazinecarbodithioate derivatives for future use as promising leads for new antidermatophyte agents development. [ABSTRACT FROM AUTHOR]

Iron is an essential nutrient for all organisms. For pathogenic fungi, iron is essential for the success of infection. Thus, these organisms have developed high affinity iron uptake mechanisms to deal with metal deprivation imposed by the host. Siderophore production is one of the mechanisms that fungal pathogens employ for iron acquisition. Paracoccidioides spp. present orthologous genes encoding the enzymes necessary for the biosynthesis of hydroxamates, and plasma membrane proteins related to the transport of these molecules. All these genes are induced in iron deprivation. In addition, it has been observed that Paracoccidioides spp. are able to use siderophores to scavenge iron. Here we observed that addition of the xenosiderophore ferrioxamine B FOB) to P. brasiliensis culture medium results in repression (at RNA and protein levels) of the SidA, the first enzyme of the siderophore biosynthesis pathway. Furthermore, SidA activity was reduced in the presence of FOB, suggesting that P. brasiliensis blocks siderophores biosynthesis and can explore siderophores in the environment to scavenge iron. In order to support the importance of siderophores on Paracoccidioides sp. life and infection cycle, silenced mutants for the sidA gene were obtained by antisense RNA technology. The obtained AsSidA strains displayed decreased siderophore biosynthesis in iron deprivation conditions and reduced virulence to an invertebrate model. [ABSTRACT FROM AUTHOR]